公开(公告)号：KR1020030035266A

公开(公告)日：2003-05-09

申请号：KR1020010067249

申请日：2001-10-30

Applicant: (주)코아바이오텍 ,  신풍제약주식회사

Inventor： 김현희 ,  유광현 ,  권장성 ,  오영애 ,  신성호

IPC: C12N15/85

CPC classification number: C12N15/85 ,  C07K14/505 ,  C12N5/0682 ,  C12P21/02

Abstract: PURPOSE: A recombinant vector for animal cells and a method for producing erythropoietin using the same vector are provided, thereby the erythropoietin can be mass-produced and the erythropoietin produced has the same physiological activity as natural erythropoietin. CONSTITUTION: The recombinant vector for animal cells comprises a promoter of cytomegalovirus containing a binding sequence of a mutated methylated DNA binding protein wherein a low affinity binding portion is substituted by a high affinity binding portion, dhfr operably linked to the cytomegalovirus promoter and replication origin. The expression vector for animal cell to mass-produce erythropoietin comprises a promoter of cytomegalovirus containing a binding sequence of modified methylated DNA binding protein wherein a low affinity binding portion is substituted by a high affinity binding portion, dhfr operably linked to the cytomegalovirus promoter, replication origin, a gene encoding erythropoietin, and a promoter operably linked to the erythropoietin gene which is operated in eukaryotic cells.

Abstract translation: 目的：提供用于动物细胞的重组载体和使用相同载体产生促红细胞生成素的方法，从而大量产生红细胞生成素，并且所产生的促红细胞生成素具有与天然促红细胞生成素相同的生理活性。 构成：用于动物细胞的重组载体包含巨细胞病毒的启动子，其含有突变的甲基化DNA结合蛋白的结合序列，其中低亲和力结合部分被高亲和力结合部分取代，dhfr可操作地连接到巨细胞病毒启动子和复制起点。 用于大规模生产促红细胞生成素的动物细胞的表达载体包含含有修饰的甲基化DNA结合蛋白的结合序列的巨细胞病毒的启动子，其中低亲和力结合部分被高亲和力结合部分取代，dhfr可操作地连接到巨细胞病毒启动子，复制 起源，编码促红细胞生成素的基因，以及与在真核细胞中操作的红细胞生成素基因可操作地连接的启动子。

公开(公告)号：KR100493703B1

公开(公告)日：2005-06-02

申请号：KR1020010067249

申请日：2001-10-30

Applicant: (주)코아바이오텍 ,  신풍제약주식회사

Inventor： 김현희 ,  유광현 ,  권장성 ,  오영애 ,  신성호

IPC: C12N15/85

Abstract: 본 발명은 벡터 증폭이 우수하게 달성되도록 재조합된 벡터 및 상기 벡터를 이용한 에리트로포이에틴의 제조방법에 관한 것으로서, 보다 상세하게는 메틸화 DNA 결합 단백질의 결합 서열 중 저친화성 결합 부위를 고친화성 결합 부위로 변이된 메틸화 DNA 결합 단백질의 결합 서열을 포함하는 사이토메갈로바이러스의 프로모터 및 상기 프로모터에 작동적으로 연결된 dhfr 및 진핵세포에서 작동하는 복제원점을 포함하는 dhfr
- 동물세포용 벡터 및 상기 벡터에 에리트로포이에틴을 코딩하는 유전자가 삽입된 발현 벡터, 그리고 그를 이용한 에리트로포이에틴의 제조방법에 관한 것이다.

公开(公告)号：KR1020030012321A

公开(公告)日：2003-02-12

申请号：KR1020010046304

申请日：2001-07-31

Applicant: (주)코아바이오텍

Inventor： 김현희 ,  유광현 ,  최완규 ,  오영애 ,  권장성 ,  신성호

IPC: C12N15/86

CPC classification number: C12N15/86 ,  C07K14/005 ,  C12N5/0682 ,  C12N2770/24222 ,  C12P21/00

Abstract: PURPOSE: Provided are expression vectors useful in animal cells and expression vectors for preparing HBV surface antigens massively, thereby mass-producing HBV surface antigen by low concentration of DHFR inhibitor. CONSTITUTION: The expression vector useful in dhfr¬-animal cells includes the promotor of Cytomegalovirus where an enhancer region is removed, dhfr linked to the promotor, and a replication origin working in a eukaryote. The expression vector useful in dhfr¬-animal cells for mass-production of HBV surface antigens contains the promotor of Cytomegalovirus where an enhancer region is removed, dhfr linked to the promotor, a replication origin working in a eukaryote, HBV surface antigen coding gene and the promotor linked to the HBV surface antigen coding gene. Wherein the eukaryote is a CHO cell.

Abstract translation: 目的：提供用于大规模制备HBV表面抗原的动物细胞和表达载体的表达载体，从而通过低浓度的DHFR抑制剂大量生产HBV表面抗原。 构成：用于dhfr-动物细胞的表达载体包括巨细胞病毒的启动子，其中去除增强子区，dhfr连接到启动子，以及在真核生物中起作用的复制起点。 用于大规模生产HBV表面抗原的动物细胞中的表达载体包含巨细胞病毒的启动子，其中除去增强子区，dhfr连接到启动子，在真核生物中起作用的复制起点，HBV表面抗原编码基因和 该启动子与HBV表面抗原编码基因相连。 其中真核生物是CHO细胞。

公开(公告)号：KR1020030078115A

公开(公告)日：2003-10-08

申请号：KR1020020016942

申请日：2002-03-28

Applicant: (주)코아바이오텍

Inventor： 신성호 ,  김현희 ,  유광현 ,  오영애 ,  권장성 ,  이우성

IPC: C07K7/08

CPC classification number: C07K7/08 ,  A61K47/34

Abstract: PURPOSE: A peptide gene carrier and a method for transfecting a gene into a cell using the same are provided, thereby stably transfecting the gene into the cell without cell toxicity. CONSTITUTION: A peptide gene carrier comprises a plurality of arginine residues, wherein the number of arginine is 5 to 14, preferably 12. A method for transfecting a gene into a cell using the peptide gene carrier comprises the steps of: (a) preparing a mixture of peptide gene carrier comprising a plurality of arginine residues, polynucleotide to be transfected, and a binding agent capable of binding the peptide gene carrier with the polynucleotide; and (b) applying the mixture to the cell, wherein the binding agent is a buffer solution having the pKa value of 6.5 to 8.5; the binding agent is phosphate buffer solution, serum free cell cultured medium, HEPES buffer solution, and MOPS buffer solution; and the cell is either normal animal cell or tumor animal cell.

Abstract translation: 目的：提供肽基因载体和使用该基因载体将基因转染入细胞的方法，从而将基因稳定转染到细胞中而没有细胞毒性。 构成：肽基因载体包含多个精氨酸残基，其中精氨酸的数目为5〜14，优选为12.使用肽基因载体将基因转染入细胞的方法包括以下步骤：（a）制备 包含多个精氨酸残基的肽基因载体的混合物，待转染的多核苷酸和能够与多核苷酸结合肽基因载体的结合剂; 和（b）将所述混合物施加到所述电池中，其中所述粘合剂是pKa值为6.5至8.5的缓冲溶液; 结合剂为磷酸盐缓冲溶液，无血清培养基，HEPES缓冲溶液和MOPS缓冲液; 细胞是正常动物细胞或肿瘤动物细胞。